Establishment and transcriptomic characterization of canine organoids from multiple tissues

Area of Science:

• Stem cell research
• Comparative medicine
• Organoid technology

Background:

• Traditional 2D cell cultures and murine models have limitations in mimicking human physiology and disease.
• Canine models offer advantages due to similarities with humans, but canine organoids are underdeveloped.
• There is a need for robust canine organoid models for drug testing and disease modeling.

Purpose of the Study:

• To establish and characterize adult-stem cell-derived canine organoid cell lines.
• To create a platform for personalized medicine and drug testing using canine organoids.
• To enable reverse translational research bridging veterinary and human medicine.

Main Methods:

• Established six canine organoid cell lines from adult stem cells (endometrium, pancreas, urinary bladder, kidney, lung, liver).
• Performed molecular characterization using immunohistochemistry and bulk RNA-sequencing (RNA-seq).
• Utilized single-cell RNA-sequencing (scRNA-seq) to identify transcriptomic signatures and cell subtypes.

Main Results:

• Successfully established and characterized six canine organoid lines from two genetically related donors.
• Bulk RNA-seq showed tissue-specific transcriptomic profiles, with organoids enriched in proliferation genes and tissues in inflammation genes.
• scRNA-seq revealed diverse epithelial subtypes and organ-based clustering.

Conclusions:

• Canine organoids provide a platform for reverse translational research, reducing animal testing.
• Genetically related donors highlight tissue-specific variations, enabling personalized medicine applications.
• These models can advance disease modeling and pharmacology in both veterinary and human research.